Chelberg M K, Tsilibary E C, Hauser A R, McCarthy J B
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455.
Cancer Res. 1989 Sep 1;49(17):4796-802.
Tumor cell metastasis involves a complex series of interdependent events, including repeated invasion of basement membranes. Studies from several laboratories have implicated tumor cell adhesion and migration in response to laminin as a major contributing factor in tumor cell invasion. The current studies address the direct role of type IV collagen in promoting tumor cell adhesion, spreading, and migration. The observations of type IV collagen-mediated cellular behavior are contrasted with cellular behavior on type I collagen. The highly metastatic K1735 M4 melanoma cell line adhered, spread, and migrated in response to type IV collagen in a concentration-dependent manner. Functional assays using well-defined proteolytic fragments of type IV collagen demonstrated that melanoma cells interact with multiple domains of this protein. Highly metastatic melanoma cells adhered, spread, and exhibited motile behavior in response to 0.2 to 200 nM concentrations of a purified pepsin-generated, triple helix-rich domain of type IV collagen. In contrast, cells adhered and spread but were essentially nonmotile in response to a purified major noncollagenous domain of the protein. In addition, de novo protein synthesis was required for cell adhesion to the major noncollagenous domain, whereas adhesion to the helical domain was less dependent upon de novo protein synthesis. Arg-Gly-Asp (RGD)-related peptides were used to study the adhesion and spreading of melanoma cells on type IV collagen. The results demonstrated that a serine containing RGD-related peptide (GRGDSP) has virtually no effect on melanoma cell adhesion on type IV collagen-coated substrata, whereas this peptide inhibited melanoma cell adhesion to fibronectin-coated substrata in a concentration-dependent manner. In contrast, when threonine was substituted for serine (GRGDTP), cell adhesion to type IV collagen was significantly (45%) inhibited. The threonine-containing peptide virtually eliminated cell adhesion on substrata coated with type I collagen. These data demonstrate that adhesion, spreading, and migration of melanoma cells on type IV collagen have a complex molecular basis which is partially dependent on RGD-related sequences.
肿瘤细胞转移涉及一系列复杂的相互依存事件，包括对基底膜的反复侵袭。多个实验室的研究表明，肿瘤细胞对层粘连蛋白的黏附与迁移是肿瘤细胞侵袭的主要促成因素。当前研究探讨了IV型胶原在促进肿瘤细胞黏附、铺展和迁移中的直接作用。将IV型胶原介导的细胞行为观察结果与I型胶原上的细胞行为进行了对比。高转移性的K1735 M4黑色素瘤细胞系对IV型胶原以浓度依赖的方式发生黏附、铺展和迁移。使用IV型胶原明确的蛋白水解片段进行的功能测定表明，黑色素瘤细胞与该蛋白的多个结构域相互作用。高转移性黑色素瘤细胞对0.2至200 nM浓度的纯化胃蛋白酶产生的富含三螺旋结构域的IV型胶原发生黏附、铺展并表现出运动行为。相比之下，细胞对该蛋白的纯化主要非胶原结构域发生黏附与铺展，但基本不运动。此外，细胞黏附于主要非胶原结构域需要从头合成蛋白质，而黏附于螺旋结构域对从头合成蛋白质的依赖性较小。使用精氨酸-甘氨酸-天冬氨酸（RGD）相关肽来研究黑色素瘤细胞在IV型胶原上的黏附与铺展。结果表明，含丝氨酸的RGD相关肽（GRGDSP）对IV型胶原包被基质上的黑色素瘤细胞黏附几乎没有影响，而该肽以浓度依赖的方式抑制黑色素瘤细胞对纤连蛋白包被基质的黏附。相比之下，当苏氨酸取代丝氨酸（GRGDTP）时，细胞对IV型胶原的黏附受到显著（45%）抑制。含苏氨酸的肽几乎消除了细胞在I型胶原包被基质上的黏附。这些数据表明，黑色素瘤细胞在IV型胶原上的黏附、铺展和迁移具有复杂的分子基础，部分依赖于RGD相关序列。
